Cell culture substrates and methods of use

C - Chemistry – Metallurgy – 12 – M

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C12M 3/04 (2006.01) A61L 31/00 (2006.01) C07K 14/78 (2006.01) C07K 17/02 (2006.01) C08H 1/06 (2006.01) C12N 5/00 (2006.01) C12N 5/06 (2006.01) C12N 11/02 (2006.01)

Patent

CA 2146946

Cell culture substrates comprising dried films of native fibrillar collagen produced by a method in which collagen fibers are hydrolyzed in acid, solubilized, and reformed as gels on porous surfaces under non-physiologic salt conditions to produce large fibers with the striations characteristic of collagen fibers found in vivo. The gels are collapsed onto the porous surfaces by drawing the interfibril fluid out of the gel through the underside of the porous surface and then dried to form films. Dried collagen films made in this manner retain native fibrillar collagen structure and excellent diffusion characteristics. Native fibrillar collagen films produced according to the methods of the invention are useful as cell culture substrates. They have particularly advantageous properties for growth and differentiation of epithelial cells. This effect is synergistically enhanced by addition of butyric acid as a differentiation inducing agent.

Des substrats pour culture cellulaire composés de pellicules sèches de collagène fibrillé indigène produites grâce à une méthode où les fibres de collagène sont hydrolysés dans l'acide, rendus plus solubles et reformés en gel sur des surfaces poreuses, dans des conditions de soluté non physiologique, pour produire de larges fibres possédant les caractéristiques de striation des fibres de collagène que l'on retrouve in vivo. Les gels tombent sur les surfaces poreuses en laissant passer le fluide interfibrillé par le dessous de la surface poreuse et sont ensuite séchés pour former les pellicules. Les pellicules de collagène séchées produites de cette façon conservent la structure du collagène fibrillé indigène ainsi que d'excellentes caractéristiques de diffusion. Les pellicules de collagène fibrillé indigène produites selon les méthodes de cette invention sont utiles en tant que substrats pour culture cellulaire. Elles possèdent des propriétés particulièrement avantageuses pour la croissance et la différenciation de cellules épithéliales. Cet effet est amélioré de façon synergétique par l'addition d'acide butyrique comme agent provoquant la différenciation.

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